A model organism for new gene discovery by cDNA sequencing
- Univ. of Iowa, Iowa City, IA (United States); and others
One method of new gene discovery is single pass sequencing of cDNAs to identify expressed sequence tags (ESTs). Model organisms can have biological properties which makes their use advantageous over studies with humans. One such model organism with advantages for cDNA sequencing is the African trypanosome T. brucei rhodesiense. This organism has the same 40 nucleotide sequence (splice leader sequence) on the 5{prime} end of all mRNAs. We have constructed a 5{prime} cDNA library by priming off the splice leader sequence and have begun sequencing this cDNA library. To date, over nearly 500 such cDNA expressed sequence tags (ESTs) have been examined. Forty-three percent of the sequences sampled from the trypanosome cDNA library have significant similarities to sequences already in the protein and translated nucleic acid databases. Among these are cDNA sequences which encode previously reported T. brucej proteins such as the VSG, tubulin, calflagin, etc., and proteins previously identified in other trypanosomatids. Other cDNAs display significant similarities to genes in unrelated organisms encoding several ribosomal proteins, metabolic enzymes, GTP binding proteins, transcription factors, cyclophillin, nucleosomal histones, histone H1, and a macrophage stress protein, among others. The 57% of the cDNAs that are not similar to sequences currently in the databases likely encode both trypanosome-specific proteins and housekeeping proteins shared with other eukaryotes. These cDNA ESTs provide new avenues of research for exploring both the biochemistry and the genome organization of this parasite, as well as a resource for identifying the 5{prime} sequence of novel genes likely to have homology to genes expressed in other organisms.
- OSTI ID:
- 134404
- Report Number(s):
- CONF-941009-; ISSN 0002-9297; TRN: 95:005313-1138
- Journal Information:
- American Journal of Human Genetics, Vol. 55, Issue Suppl.3; Conference: 44. annual meeting of the American Society of Human Genetics, Montreal (Canada), 18-22 Oct 1994; Other Information: PBD: Sep 1994
- Country of Publication:
- United States
- Language:
- English
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